LOCUS       sequence-128084-        4505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TOPO construct expressing iRFP670 for generation of GMAP-compatible 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4505)
  AUTHORS   Akama-Garren EH, Joshi NS, Tammela T, Chang GP, Wagner BL, Lee DY, 
            Rideout Iii WM, Papagiannakopoulos T, Xue W, Jacks T
  TITLE     A Modular Assembly Platform for Rapid Generation of DNA Constructs.
  JOURNAL   Sci Rep. 2016 Feb 18;6:16836. doi: 10.1038/srep16836.
  PUBMED    26887506
REFERENCE   2  (bases 1 to 4505)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-02-18"; volume: "6:16836. doi"; pages: " 
            10.1038/srep16836"
COMMENT     Sequence Label: TOPO iRFP670
FEATURES             Location/Qualifiers
     source          1..4505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1392..1411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1393..1411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1418..1435)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1418..1434)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1427..1449)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             1572..1874
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1753..1772
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             2223..3017
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2223..3017
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2277..2296)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2887..2906
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      3218..3227
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3224..3598
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      3736..4324
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4225..4244
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4478..4495
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcct
      301 aactcgaacg ctagctgtgc gatcgtttac catggcacgg aaggtagatc tcacgagttg
      361 tgatagagaa ccaatacata tcccggggag catccaacca tgtggttgct tgctcgcatg
      421 cgacgcacag gctgtaagaa taactcggat caccgaaaat gcaggggcct ttttcggtag
      481 agaaaccccg agagttgggg agctgctggc cgattatttt ggagagactg aggcacatgc
      541 attgcgaaac gcactggccc agtcaagcga ccctaagcga ccagctctta tcttcggctg
      601 gcgcgatggc ttgactggcc gcactttcga tatctccctg cacagacacg atggtacatc
      661 tatcattgag ttcgaaccgg ccgcagctga acaggccgat aacccattga gattgaccag
      721 gcagatcatc gccaggacga aagaattgaa gtcactggag gagatggcag caagagtccc
      781 taggtatttg caagcaatgc tgggatacca ccgcgttatg ctttaccgat ttgcagacga
      841 cgggtccggt atggtgatag gagaggccaa gcgctccgac ctggagtcat ttcttggcca
      901 acactttcct gccagcctgg taccccagca agctaggttg ctctatctca aaaatgctat
      961 acgggtggtg agcgactcca ggggcattag cagccgcatc gtgcccgagc acgacgctag
     1021 tggtgccgcc ctcgaccttt cctttgctca tctccgaagc atctctccat gccacctgga
     1081 attcctgaga aacatgggtg tgtcagcttc catgagcctg tccatcatca tcgacggaac
     1141 tctctgggga ctgattatct gtcatcatta tgaacctcgc gctgtcccca tggcccaacg
     1201 cgtggcagct gaaatgtttg ccgacttcct gtctcttcac ttcactgccg ctcatcacca
     1261 gaggtgaaaa cgttgttgtt tggggttgaa ttactctgga tccactagta acggccgcca
     1321 gtgtgctgga attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag
     1381 ggcccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     1441 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     1501 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1561 tatacgtacg gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg
     1621 tggatgtaca gagtgatatt attgacacgc cggggcgacg gatggtgatc cccctggcca
     1681 gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg
     1741 atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg
     1801 aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt
     1861 tctggggaat ataaatgtca ggcatgagat tatcaaaaag gatcttcacc tagatccttt
     1921 tcacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg
     1981 ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta
     2041 catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg
     2101 gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttctcgccgc
     2161 caaggatctg atggcgcagg ggatcaagct ctgatcaaga gacaggatga ggatcgtttc
     2221 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     2281 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     2341 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     2401 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     2461 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     2521 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     2581 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     2641 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     2701 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     2761 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2821 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     2881 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2941 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3001 ttgacgagtt cttctgaatt attaacgctt acaatttcct gatgcggtat tttctcctta
     3061 cgcatctgtg cggtatttca caccgcatac aggtggcact tttcggggaa atgtgcgcgg
     3121 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3181 accctgataa atgcttcaat aatagcacgt gaggagggcc accatggcca agttgaccag
     3241 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     3301 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     3361 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     3421 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     3481 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     3541 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     3601 cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3661 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3721 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3781 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3841 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3901 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3961 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4021 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4081 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4141 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4201 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4261 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4321 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4381 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4441 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4501 ggaag
//
